CN114058543B - Pediococcus acidilactici DY15, application thereof, feed and preparation method thereof - Google Patents
Pediococcus acidilactici DY15, application thereof, feed and preparation method thereof Download PDFInfo
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- CN114058543B CN114058543B CN202111384226.2A CN202111384226A CN114058543B CN 114058543 B CN114058543 B CN 114058543B CN 202111384226 A CN202111384226 A CN 202111384226A CN 114058543 B CN114058543 B CN 114058543B
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- pediococcus acidilactici
- feed
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- fermentation
- preparation
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Classifications
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- A—HUMAN NECESSITIES
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- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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Abstract
The invention provides pediococcus acidilactici DY15, application thereof, feed and a preparation method thereof, and relates to the technical field of biology. The research of the inventor shows that the Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention has antibacterial activity, has good inhibition effect on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product comprises various flavor substances, can be applied to the field of feed, improves the nutritive value of the feed, improves the palatability, improves the immunity of livestock and poultry, and establishes a good intestinal microbial system. According to the preparation method of the feed, disclosed by the invention, the palm meal and the bean skin are treated in a mode of synergistic fermentation by using the bacterial enzymes, so that the method is simple and convenient, the antibacterial capacity and the content of organic acid of the palm meal and the bean skin are greatly improved, the palatability of the feed is also improved, and the palm meal and the bean skin become possible.
Description
Technical Field
The invention relates to the technical field of biology, in particular to pediococcus acidilactici DY15, application thereof, feed and a preparation method thereof.
Background
Lactic acid bacteria are a class of gram-positive, spore-free bacteria collectively known as lactic acid, the major metabolite of which ferments sugars. Lactic acid bacteria, which are a living microorganism and play an important role in human and animal health, have been used in various fields such as food fermentation, livestock industry, medical care and industrial production of lactic acid, and thus have become an important object of research. Wherein the health effect of Pediococcus acidilactici is also used for treating intestinal infection and inflammatory bowel disease, and the addition of lactobacillus in poultry and livestock industry has the effect of antibiotics instead of antibiotics, thus enriching the nutrition of feed, maintaining the balance of intestinal flora and promoting the healthy development of animals.
The palm meal is a byproduct of oil extraction of oil palm fruits, has the characteristics of huge yield, low price, no aflatoxin, and high content of fat, crude fiber and amino acid. The soybean hulls are byproducts of soybean processing, have low lignin content and high fiber content, and are easy to absorb. The maximum adding proportion of the daily ration cannot be accurately determined when the daily ration is prepared due to the limitation of nutritional ingredients and feeding value evaluation of unconventional feed raw materials. In addition, as the total carbohydrate in the palm meal is mostly non-starch polysaccharide such as mannan which is difficult to digest, the soybean hulls also have anti-nutritional factors such as glycinin which influence the digestion and absorption of nutritional ingredients, and the content of polypeptide substances is low, so that the digestion and absorption of animals are not facilitated.
In view of this, the present invention has been made.
Disclosure of Invention
The first object of the present invention is to provide a pediococcus acidilactici (Pediococcus acidilactici) DY15 deposited with the chinese collection of typical cultures at 10 and 18 of 2021 under the deposit number: cctccc NO: m20211291.
The second object of the present invention is to provide a microbial agent.
The third object of the invention is to provide the application of the pediococcus acidilactici (Pediococcus acidilactici) DY15 or the microbial inoculum in preparing antibacterial products.
The fourth object of the invention is to provide the application of the pediococcus acidilactici (Pediococcus acidilactici) DY15 or the microbial inoculum in feed.
A fifth object of the present invention is to provide a method for preparing a feed.
A sixth object of the present invention is to provide a feed.
In a first aspect, the present invention provides a pediococcus acidilactici (Pediococcus acidilactici) DY15, wherein the pediococcus acidilactici (Pediococcus acidilactici) DY15 is deposited with the chinese collection of typical cultures, with deposit numbers: cctccc NO: m20211291.
In a second aspect, the invention provides a microbial inoculum comprising Pediococcus acidilactici (Pediococcus acidilactici) DY15 as described above.
In a third aspect, the invention provides an application of the pediococcus acidilactici (Pediococcus acidilactici) DY15 or the microbial inoculum in preparing antibacterial products.
As a further technical scheme, the bacteriostasis comprises inhibiting at least one of staphylococcus aureus, salmonella or escherichia coli.
In a fourth aspect, the invention provides the use of Pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum as described above in any one of the following a to c:
a. preparing feed;
b. improving the palatability of the feed;
c. improving the nutritive value of the feed.
In a fifth aspect, the invention provides a method for preparing feed, comprising the steps of: mixing a substrate with alkaline protease, mannanase and the pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum, and fermenting to prepare a feed;
the substrate comprises palm meal and bean curd skin.
As a further technical scheme, the mass ratio of the palm meal to the bean curd skin is 6:2-6:6, preferably 6:4;
preferably, the substrate further comprises molasses; the mass percentage of molasses in the substrate is 1% -5%, preferably 3%.
As a further technical scheme, after mixing, the concentration of the pediococcus acidilactici (Pediococcus acidilactici) DY15 is 1 multiplied by 10 7 CFU/g or more;
preferably, the alkaline protease is used in an amount of 1000 to 1500U/g substrate, preferably 1200U/g substrate;
the amount of mannanase is 2000-3000U/g substrate, preferably 2500U/g substrate.
As a further technical scheme, the water content of the substrate is 45-65%, preferably 55%;
preferably, the temperature of the fermentation is 36-38 ℃, preferably 37 ℃;
preferably, the fermentation time is 36-60 hours, preferably 48 hours;
preferably, the fermentation is performed in a sealed manner.
In a sixth aspect, the invention provides a feed prepared by the preparation method.
Compared with the prior art, the invention has the following beneficial effects:
the research of the inventor shows that the Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention has antibacterial activity, has good inhibition effect on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product comprises various flavor substances, can be applied to the field of feed, improves the nutritive value of the feed, improves the palatability, improves the immunity of livestock and poultry, and establishes a good intestinal microbial system.
The preparation method of the feed provided by the invention takes palm meal and soybean hulls as substrates, takes pediococcus acidilactici (Pediococcus acidilactici) DY15 as zymophyte, simultaneously adds alkaline protease and mannanase, and adopts a mode of synergistic fermentation of bacterial enzymes to treat the palm meal and the soybean hulls; after fermentation, the lactic acid content is increased by 671.93 times compared with that before fermentation, the total acid content is increased by 6.34 times compared with that before fermentation, the acid production capacity is moderate, and excessive acidification of palm meal and beans is not caused; the content of the small peptide is increased by 28.03% compared with the content before fermentation, the added value of the raw materials is increased, and the waste recycling is facilitated; after fermentation, the substances with different contents such as ethyl octanoate, methyl laurate, acetoin, methyl myristate, acetic acid, isoamyl octanoate, phenylacetaldehyde and the like are detected, and many substances are common materials for preparing essence and spice, so that the palatability of the fermented palm meal and bean skin is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the morphology of Pediococcus acidilactici DY15 in a plate medium;
FIG. 2 shows the growth curve of Pediococcus acidilactici DY15 in MRS medium.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to embodiments and examples, but it will be understood by those skilled in the art that the following embodiments and examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The specific conditions are not specified, and the process is carried out according to conventional conditions or conditions suggested by manufacturers. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
In a first aspect, the invention provides a strain of Pediococcus acidilactici (Pediococcus acidilactici) DY15, the strain being classified as: pediococcus acidilactici DY15, latin Wen Xueming: pediococcus acidilactici DY15, deposited at China center for type culture Collection, accession number: chinese, wuhan, university of Wuhan, date of preservation: 2021, 10 months and 18 days, deposit number: cctccc NO: m20211291.
The Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention is separated from tin-free European Shang Chaoshi pickled Chinese cabbage, the bacterial cells are of medium size, the bacterial colony on the culture medium is milky white, the surface is smooth and moist, and the bacterial colony is raised, so that the Pediococcus acidilactici has a peculiar sour and fragrant taste.
The research of the inventor shows that the Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention has antibacterial activity, has good inhibition effect on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product comprises various flavor substances, can be applied to the field of feed, improves the nutritive value of the feed, improves the palatability, improves the immunity of livestock and poultry, and establishes a good intestinal microbial system.
In a second aspect, the present invention provides a microbial inoculum comprising Pediococcus acidilactici (Pediococcus acidilactici) DY15 as described above, or further comprising other strains or adjuvants known to those skilled in the art, which has all the beneficial effects of the strain of the present invention, as it contains Pediococcus acidilactici (Pediococcus acidilactici) DY15.
In a third aspect, the invention provides an application of the pediococcus acidilactici (Pediococcus acidilactici) DY15 or the microbial inoculum in preparing antibacterial products.
The research of the inventor shows that the Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention has antibacterial activity and good inhibition effect on the growth of escherichia coli, salmonella and staphylococcus aureus, so that the Pediococcus acidilactici can be used for preparing antibacterial products.
In a fourth aspect, the invention provides the use of Pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum as described above in any one of the following a to c:
a. preparing feed;
b. improving the palatability of the feed;
c. improving the nutritive value of the feed.
The research of the inventor shows that the Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention has antibacterial activity, has good inhibition effect on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product comprises various flavor substances, can be applied to the field of feed, improves the nutritive value of the feed, improves the palatability, improves the immunity of livestock and poultry, and establishes a good intestinal microbial system. Therefore, the Pediococcus acidilactici (Pediococcus acidilactici) DY15 provided by the invention can be applied to preparing feed, improving the palatability of the feed and improving the nutritional value of the feed.
In a fifth aspect, the invention provides a method for preparing feed, comprising the steps of: mixing a substrate with alkaline protease, mannanase and the pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum, and fermenting to prepare a feed;
the substrate comprises palm meal and bean curd skin.
The preparation method provided by the invention is simple and convenient, adopts a mode of synergistic fermentation of bacterial enzymes to treat the palm meal and the bean skin, improves the bacteriostatic ability and the content of organic acid of the palm meal and the bean skin, can prevent the field planting of pathogenic bacteria in animal intestinal tracts, enhances the disease resistance of animals, reduces anti-nutritional factors in raw materials, improves the utilization rate of nutritional substances in feed and the digestion ability of animals, and has great significance for improving the feeding value of the palm meal and the bean skin and the utilization of unconventional feed.
As a further technical solution, the mass ratio of the palm meal to the beancurd sheet may be, for example, but not limited to, 6:2, 6:3, 6:4, 6:5 or 6:6, preferably 6:4;
preferably, the substrate further comprises molasses; the mass percentage of molasses in the substrate may be, for example, but not limited to, 1%, 2%, 3%, 4% or 5%, preferably 3%.
As a further technical scheme, after mixing, the concentration of the pediococcus acidilactici (Pediococcus acidilactici) DY15 is 1 multiplied by 10 7 CFU/g or more;
preferably, the alkaline protease may be used in an amount of, for example, but not limited to, 1000U/g substrate, 1100U/g substrate, 1200U/g substrate, 1300U/g substrate, 1400U/g substrate or 1500U/g substrate, preferably 1200U/g substrate;
the mannanase may be used in an amount of, for example but not limited to, 2000U/g substrate, 2200U/g substrate, 2400U/g substrate, 2600U/g substrate, 2800U/g substrate or 3000U/g substrate, preferably 2500U/g substrate.
"1200U/g substrate" means that 1g substrate was added to 1200U of the enzyme.
As a further embodiment, the substrate may comprise, for example, but not limited to, 45%, 49%, 53%, 57%, 61% or 65%, preferably 55% by weight;
preferably, the temperature of the fermentation may be, for example, but not limited to, 36 ℃,37 ℃ or 38 ℃, preferably 37 ℃;
preferably, the fermentation time can be, for example, but not limited to, 36h, 40h, 44h, 48h, 52h, 56h or 60h, preferably 48h;
preferably, the fermentation is performed in a sealed manner.
By further optimizing and adjusting the fermentation process, the fermentation of the palm meal and the bean skin is better realized, and the palatability and the nutritional value of the feed are improved.
In a sixth aspect, the invention provides a feed prepared by the preparation method.
The feed provided by the invention has high nutritive value, is easy to digest and good in palatability, and can enhance the disease resistance of livestock.
The invention is further illustrated by the following specific examples and comparative examples, however, it should be understood that these examples are for the purpose of illustration only in greater detail and should not be construed as limiting the invention in any way.
Example 1 screening of strains
1 culture medium: MRS Medium (g/L): 10.0 parts of peptone, 8.0 parts of beef extract, 4.0 parts of yeast powder, 20.0 parts of glucose, 2.0 parts of dipotassium hydrogen phosphate, 2.0 parts of triammonium citrate, 5.0 parts of sodium acetate, 0.58 parts of magnesium sulfate heptahydrate, 0.25 parts of manganese sulfate tetrahydrate, 1mL of Tween 80, 1L of distilled water and 20 minutes of sterilization at 115 ℃.
2, screening: the strain is separated from tin-free Europe Shang Chaoshi, pickled Chinese cabbage is firstly placed in an MRS culture medium for culturing for 24 hours at 37 ℃, the culture solution is respectively diluted by 100 and 1000 times by sterile water, 100 mu L of the culture solution is coated on the MRS solid culture medium for culturing for 24 hours at 37 ℃, and a lactic acid strain is obtained by streaking separation (shown in figure 1). The microbiological characteristics are that gram staining is typically positive, bacterial colony is medium in size, bacterial colony on the culture medium is milky white, the surface is smooth and moist, and the microbial characteristic is that the microbial characteristic has peculiar sour and fragrant taste. The strain was cultured in MRS medium and its growth curve is shown in FIG. 2.
3: identification of strains: the screened strain is coated on an MRS plate, single colony is picked and amplified by using a general primer 1492R (GGTTACCTTGTTACGACTT (SEQ ID NO. 1)) and 27F (AGAGTTTGATCCTGGCTCAG (SEQ ID NO. 2)), the amplified product is delivered to a biological engineering (Shanghai) limited company for 16SrRNA sequencing, and the sequence obtained results are subjected to homology comparison with model strains in Genbank through NCBI's Nucleoteide BLAST. The comparison shows 99% similarity with 16sRNA of the related model strain in Genbank, and the strain is determined to be Pediococcus acidilactici and named Pediococcus acidilactici (Pediococcus acidilactici) DY15.
Example 2 identification of physiological and Biochemical characteristics of Pediococcus acidilactici DY15
(1) Litmus milk experiment: milk mainly contains lactose and casein, and is added with litmus as an indicator and a redox indicator, and the litmus is discolored from top to bottom to be reduced to white when the litmus is in pink alkaline and reduced to blue when the litmus is in light purple acidic and neutral. Lactic acid bacteria ferment lactose to produce acid, and litmus turns red, and when the acidity is high, milk can be coagulated. And observing the experimental results of the inoculated and cultured litmus milk, and observing the acidogenesis and coagulation reaction of the litmus milk after one day of culture at 37 ℃.
(2) Starch hydrolysis experiments: inoculating the strain into basic culture medium containing soluble starch, culturing at 37deg.C for 24 hr, collecting a little culture solution in colorimetric disc, collecting non-inoculated culture solution as control, and adding Lu Geshi iodine solution respectively. Non-chromogenic indicates starch hydrolysis, and bluish black or bluish purple indicates starch not hydrolyzed or not hydrolyzed completely.
(3) Gelatin liquefaction experiments: the experimental strain was inoculated in basal medium containing gelatin, incubated at 37℃for 24 hours, and an unvaccinated test tube was used as a control. The inoculated and unvaccinated control tubes were placed in a refrigerator at 4℃and after waiting for the control tube to solidify, the experimental results were recorded and the comparison was repeated several times. If the control tube solidifies, the inoculation tube liquefies to a positive reaction and the solidification to a negative reaction.
(4) V-P experiment: inoculating the experimental strain into PYG culture medium, culturing at 37deg.C for 24 hr, adding tea phenol and potassium hydroxide into the culture solution, mixing in uncapped test tube, and sequentially shaking for 30 min to show red positive reaction.
(5) Dextran experiments: the fresh culture was inoculated on a slant of a sucrose-containing medium and incubated at 37℃for 24 hours. The cultures formed a thick lawn on the slants, indicating that dextran was produced, a positive reaction, and a negative reaction.
(6) Hydrogen sulfide experiments: inoculating fresh culture into cysteine or cystine-containing culture medium, holding lead acetate filter paper with sterile forceps, suspending in inoculating test tube, closing the lower end of the test tube near the surface of the culture medium without contacting the liquid surface, tightly sealing the upper end with cotton plug, setting blank control in experiment, suspending lead acetate paper on the non-inoculated test tube culture medium, culturing at 37deg.C for 24 hr, and observing and comparing to obtain black paper as positive reaction.
(7) Sugar fermentation experiment: carbohydrates such as sugar or alcohols to be measured are added to the basal medium, and the mixture is separately packed into 5mL test tubes. Shaking culture was performed at 37℃for 24 hours. When in detection, a little culture solution is placed in a color comparison disc, meanwhile, the culture solution without carbohydrate is taken as a control, BTB-MR reagent is dripped to compare the color change, and the acid production intensity is recorded.
The physiological and biochemical characteristics of DY15 strain are shown in Table 1 and Table 2, and the results show that the litmus milk and V-P experiments are positive, the starch hydrolysis, the hydrogen sulfide experiments, the glucan and the gelatin liquefaction are negative, and the D-galactose, the maltose, the rhamnose, the sucrose, the fructose, the lactose, the glucose and the D-mannose can be utilized for producing acid by fermentation.
TABLE 1 physiological Properties of Pediococcus acidilactici DY15
Note that: "-" indicates negative, "+" indicates positive.
TABLE 2 fermentation results of Pediococcus acidilactici DY15 sugar
Note that: "+" indicates strong fermentation and "d" indicates slightly fermentation.
Example 3 determination of antibacterial diameter of Strain
Preparation of the indicator fungus liquid: three indicator bacteria of escherichia coli, salmonella and staphylococcus aureus are inoculated in an LB liquid medium and cultured for 24 hours at 37 ℃.
The oxford cup method is adopted: taking flat plates with the diameter of about 90mm, pouring 15-20mL of heated and melted nutrient agar culture medium respectively, uniformly spreading the nutrient agar culture medium in the flat plates, and placing the flat plates on a horizontal table top to solidify the nutrient agar culture medium to serve as a bottom layer. Heating and thawing semi-solid nutrient agar culture medium (agar content of 1%) in proper amount, cooling to 48-50deg.C, and adding 0.1-0.2mL (with fungus concentration of 10) of indicator fungus suspension per 50-100mL culture medium 8 CFU/mL), 5mL was added to each 1 plate, and the mixture was spread evenly over the bottom layer to form a fungus layer. 4 oxford cups are uniformly arranged in each 1 plate at equal intervals for standby, 200 mu L of lactobacillus supernatant is respectively dripped into each oxford cup in each double-layer plate, and after the culture is carried out for 10-13 hours at 37 ℃, the diameter (or area) of each bacteriostasis zone is measured, and the results are shown in Table 3.
TABLE 3 antibacterial effect of Pediococcus acidilactici DY15
EXAMPLE 4 preparation of Pediococcus acidilactici microbial agent
(1) Inoculating the original strain of Pediococcus acidilactici into MRS culture medium according to 1% of inoculum size, activating at 37 ℃ for 24h, activating for 2-3 generations in the same way, and culturing in MRS culture medium to obtain the bacterial liquid of Pediococcus acidilactici.
(2) Preparing lactobacillus bacterial liquid: the original pediococcus acidilactici strain is put into an MRS liquid culture medium for activation for 24 hours at 37 ℃, the activation is carried out for 2 times according to the steps, and then the fermentation broth is obtained after the fermentation broth is cultured for 24 hours at 37 ℃ of the MRS liquid culture medium.
Colony count: 1mL of the fermentation broth was diluted by 10-fold dilution with a sterilized 1.5mL centrifuge tube in an ultra clean bench. 100uL of the suspension was pipetted, poured into a centrifuge tube containing 900uL of distilled water, repeatedly blown 5 times into the liquid, and mixed by shaking (note the gun head was replaced and mixed by shaking thoroughly every dilution). Step-by-step dilution according to the above steps, taking 10 -7 ，10 -8 Two gradient bacterial suspensions (100. Mu.L) were added dropwise to the seed culture plates, plating was performed, and colonies were counted.
Example 5 Process for synergistic fermentation of palm meal and beancurd skin with bacterial enzymes
Using a coarse stalk grinder to mix raw materials of palm meal and bean skin according to a mass ratio of 6:4 is crushed to 2cm plus or minus 0.5cm for standby. Adding water, pediococcus acidilactici DY15, alkaline protease and mannanase into palm meal and bean curd skin for fermentation; the palm meal and the soybean hulls were allowed to have a moisture content of 55% (m/m), alkaline protease was allowed to have a substrate content of 1200U/g, mannanase was allowed to have a substrate content of 2500U/g, pediococcus acidilactici DY15 was cultured as in example 4, and the bacterial liquid was added to the substrate in an inoculum size of 5% (v/m, mL/g) to give a bacterial concentration of 1X 10 7 CFU/g。
The content of the small peptide is 62.72mg/g by adopting a Folin-phenol method, which is improved by 28.03%; the total acid content is 5.07% by acid-base titration, which is improved by 6.34 times; the number of lactic acid bacteria was detected to be 2X 10 7 cfu/g, 50% improvement; the organic acid content in the fermentation material obtained in example 5 was measured by an HPLC method, and as a result, as shown in Table 4, it was found that the contents of lactic acid, acetic acid, etc. were significantly improved. Wherein the content of lactic acid after fermentation is increased 671.93 times.
TABLE 4 Pediococcus acidilactici DY15 organic acid content
Note that: the control group was the raw material which had not been subjected to fermentation treatment.
Example 6: aromatic substance for improving palatability of feed before and after fermentation
The feed flavoring agent is also called as phagostimulant and appetite promoter, and its action principle is closely related to the functions of taste, smell, respiratory system, digestive system, etc. of animal, so that it can raise palatability of feed. The headspace test results show that the fermented palm meal and bean skin flavor substances obtained in the embodiment 5 have higher relative contents of ethyl octanoate, methyl laurate, acetoin, methyl myristate, acetic acid, isoamyl octanoate, phenylacetaldehyde and the like. The ethyl octoate is mainly used for preparing food flavoring agent and spice, has fragrance similar to brandy and has sweet taste; acetoin is mainly used for preparing edible flavors such as cream, dairy products, yoghourt, strawberries and the like, has strong cream and fat fragrance, and has pleasant milk fragrance after being diluted highly; the methyl laurate is mainly used for preparing edible essence and daily chemical essence, and has the fragrance of fat, flower fragrance and wine; methyl myristate is commonly used in edible essence such as honey, coconut and the like, is also used in daily essence, and is also used in organic synthesis. Malt phenol, ethyl decanoate, myristic acid, delta-dodecalactone, ethyl palmitate and the like are also detected in the fermented feed and are common materials for preparing essence and spice. In addition, it has been detected that formic acid, ammonium acetate, propionic acid, butyric acid, L-lactic acid, caproic acid and the like inhibit the growth of pathogenic bacteria and undesirable spoilage microorganisms.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
SEQUENCE LISTING
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Claims (14)
1. Pediococcus acidilacticiPediococcus acidilactici) DY15, wherein said pediococcus acidilactici (Pediococcus acidilactici) DY15 is deposited with the chinese collection of typical cultures under the deposit number: cctccc NO: m20211291.
2. A microbial inoculum, which is characterized by comprising the Pediococcus acidilactici of claim 1Pediococcus acidilactici）DY15。
3. The Pediococcus acidilactici of claim 1Pediococcus acidilactici) Use of DY15 or the microbial agent of claim 2 in the preparation of a bacteriostatic product;
the antibacterial agent is used for inhibiting at least one of staphylococcus aureus, salmonella or escherichia coli.
4. The Pediococcus acidilactici of claim 1Pediococcus acidilactici) Application of DY15 or the microbial inoculum of claim 2 in any one of the following a-c:
a. preparing feed;
b. improving the palatability of the feed;
c. improving the nutritive value of the feed.
5. The preparation method of the feed is characterized by comprising the following steps: mixing the substrate with alkaline protease, mannanase and Pediococcus acidilactici as defined in claim 1Pediococcus acidilactici) DY15 or the microbial inoculum of claim 2, and preparing feed by fermentation;
the substrate comprises palm meal and bean curd skin.
6. The preparation method of claim 5, wherein the mass ratio of the palm meal to the bean curd skin is 6:2-6:6.
7. The method according to claim 6, wherein the mass ratio of the palm meal to the bean curd skin is 6:4.
8. The method of claim 5, wherein the substrate further comprises molasses; the mass percentage of molasses in the substrate is 1% -5%.
9. The method according to claim 8, wherein the mass percentage of molasses in the substrate is 3%.
10. The method according to claim 5, wherein the concentration of DY15 in the pediococcus acidilactici (Pediococcus acidilactici) after mixing is 1X 10 7 CFU/g or more;
the consumption of the alkaline protease is 1000-1500U/g substrate;
the dosage of the mannanase is 2000-3000U/g substrate.
11. The method of claim 10, wherein the alkaline protease is used in an amount of 1200U/g substrate;
the amount of the mannanase is 2500U/g substrate.
12. The preparation method of claim 5, wherein the substrate contains 45% -65% of water by mass;
the fermentation temperature is 36-38 ℃;
the fermentation time is 36-60 hours;
the fermentation mode is sealed fermentation.
13. The method of claim 12, wherein the substrate comprises 55% water by mass;
the temperature of the fermentation is 37 ℃;
the fermentation time is 48 hours;
the fermentation mode is sealed fermentation.
14. A feed prepared by the method of any one of claims 5-13.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105410337A (en) * | 2015-12-11 | 2016-03-23 | 上海邦成生物科技有限公司 | Biological feed additive rich in bioactive peptides and probiotics as well as preparation method of additive |
| CN107034163A (en) * | 2017-05-23 | 2017-08-11 | 北京市农林科学院 | A kind of novel lactic piece coccus and its application |
| CN107227279A (en) * | 2017-07-18 | 2017-10-03 | 邓禹 | One plant of Pediococcus acidilactici and its application |
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| KR101296067B1 (en) * | 2012-11-28 | 2013-08-12 | 제이나인바이오 영농조합법인 | Health promoting foods containing pediococcus acidilactici j9 and method of manufacturing therof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105410337A (en) * | 2015-12-11 | 2016-03-23 | 上海邦成生物科技有限公司 | Biological feed additive rich in bioactive peptides and probiotics as well as preparation method of additive |
| CN107034163A (en) * | 2017-05-23 | 2017-08-11 | 北京市农林科学院 | A kind of novel lactic piece coccus and its application |
| CN107227279A (en) * | 2017-07-18 | 2017-10-03 | 邓禹 | One plant of Pediococcus acidilactici and its application |
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| Title |
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| 党建章主编.发酵工艺教程.中国轻工业出版社,2016,(第2版),145. * |
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